US2016208282A1PendingUtilityA1

Systems for cloning plants through asexual means

39
Assignee: CT DE INVESTIGACION Y ESTUDIOS AVANZADOS DEL INST POLITECNICO NAC CINVESTAVPriority: Sep 23, 2013Filed: Sep 22, 2014Published: Jul 21, 2016
Est. expirySep 23, 2033(~7.2 yrs left)· nominal 20-yr term from priority
Inventors:Jean-Philippe Vielle CalzadaCarlos Barragan RosilloEdgar Demesa ArevaloElvira Hernandez LaganaGloria Leon MartinezNidia Sanchez LeonDaniel Rodriguez LealIsaac Rodriguez ArevaloJaime Padilla Calzada
C12N 15/8218C12N 15/8261A01H 4/006C12N 15/8287A01H 1/02A01H 1/022
39
PatentIndex Score
0
Cited by
0
References
0
Claims

Abstract

Disclosed are methods of obtaining clonal seeds, methods of plant cloning, methods of screening for maternal plants that produce clonal seeds asexually and methods of increasing yield of clonal seeds. Also disclosed are constructs comprising a nucleic acid that can silence the activity of a RNA-dependent DNA methylation pathway gene. Further disclosed are maternal plants comprising a construct wherein the construct comprises an exogenous nucleic acid sequence, wherein the construct renders the maternal plant defective for RNA-dependent DNA methylation.

Claims

exact text as granted — not AI-modified
What is claimed is: 
     
         1 . A method of obtaining seeds comprising clonal embryos comprising
 collecting one or more seeds produced by a maternal plant, wherein the maternal plant is unable to be pollinated,   wherein the one or more seeds comprise a parthenogenically-derived embryo that is a clone of the maternal plant.   
     
     
         2 . The method of  claim 1 , wherein the maternal plant is defective in at least one RNA dependent DNA methylation pathway gene. 
     
     
         3 . The method of  claim 2 , wherein the RNA dependent DNA methylation pathway gene is AGO4 (ARGONAUTE 4), AGO6 (ARGONAUTE 6), AGO8 (ARGONAUTE 8), AGO9 (ARGONAUTE 9), CMT3 (CHROMOMETHYLASE 3), DCL3 (DICER-LIKE 3), DRM2 (DOMAINS REARRANGED METHYLASE 2), EXS1 (EXTRA SPOROGENOUS CELLS1), IDN2 (INVOLVED IN DE NOVO 2), MET1 (METHYL TRANSFERASE 1), NPRD1a (NUCLEAR POLYMERASE D 1a), NRPD1b (NUCLEAR POLYMERASE D 1b), NRPD2 (NUCLEAR POLYMERASE D 2), NRPE1 (NUCLEAR RNA POLYMERASE E 1), NRPE2 (NUCLEAR RNA POLYMERASE E 2), RDR2 (RNA-DEPENDENT RNA POLYMERASE 2), RDR6 (RNA-DEPENDENT RNA POLYMERASE 6), SGS3 (SUPPRESSOR OF GENE SILENCING 3), SUVH2 (SUPPRESSOR OF VARIEGATION 3-9 HOMOLOG 2), and SUVH9 (SUPPRESSOR OF VARIEGATION 3-9 HOMOLOG 9). NO OTHER GENES 
     
     
         4 . The method of  claim 3 , wherein the AGO4 allele is ago4-6 or ago4-1. 
     
     
         5 . The method of  claim 3 , wherein the AGO6 allele is ago6-2. 
     
     
         6 . The method of  claim 3 , wherein the AGO9 allele is 9-2, 9-3 or 9-4. 
     
     
         7 . The method of  claim 3 , wherein the AGO8 allele is ago 8-1. 
     
     
         8 . The method of  claim 3 , wherein the RDR2 allele is rdr2-1. 
     
     
         9 . The method of  claim 3 , wherein the RDR6 allele is rdr6-15 or rdr6-11. 
     
     
         10 . The method of  claim 3 , wherein the SGS3 allele is sgs3-11. 
     
     
         11 . The method of  claim 3 , wherein the DRM2 allele is drm2-2. 
     
     
         12 . The method of  claim 3 , wherein the MET1 allele is met1-7. 
     
     
         13 . The method of  claim 1 , wherein the maternal plant is unable to self-pollinate. 
     
     
         14 . The method of  claim 13 , wherein the maternal plant's ability to self-pollinate is disrupted physically, chemically or genetically. 
     
     
         15 . The method of  claim 14 , wherein chemical disruption comprises exposure to a gametocide that abolishes pollen formation. 
     
     
         16 . The method of  claim 15 , wherein the gametocide includes at least one of maleic hydrazide (1,2-dihydropyridazine, 3-6-dione) (MH), 2,4-dichlorophenoxyacetic acid (2,4-D), a-naphthalene acetic acid (NAA), and tri-iodobenzoic acid (TIBA). 
     
     
         17 . The method of  claim 14 , wherein physical disruption comprises emasculating the maternal plant. 
     
     
         18 . A method of obtaining seeds comprising clonal embryos comprising
 a) collecting one or more seeds produced by a maternal plant, wherein the maternal plant is defective in at least one RNA dependent DNA methylation pathway gene; and   b) sorting the seeds to separate the seeds comprising clonal embryos from the seeds comprising non-clonal embryos;   
       wherein the maternal plant is pollinated prior to collecting the seeds, and 
       wherein the one or more seeds produced by the maternal plant comprise an embryo that is a clone of the maternal plant. 
     
     
         19 . The method of  claim 18 , wherein sorting the seeds is based on phenotype or genotype. 
     
     
         20 . The method of  claim 19 , wherein the phenotype comprises size, shape, color, or a combination thereof of the seeds. 
     
     
         21 . The method of  claim 18 , wherein the sorting is performed manually. 
     
     
         22 . The method of  claim 18 , wherein the sorting is performed automatically. 
     
     
         23 . The method of  claim 22 , wherein the automatic sorting comprises a machine comprising an optical detector. 
     
     
         24 . The method of  claim 18 , wherein the sorting is done visually. 
     
     
         25 . A method of screening for maternal plants that produce seeds comprising parthenogenically-derived clonal embryos comprising
 a) obtaining a maternal plant unable to pass on paternally-derived chromosomes to embryos, wherein an activity of a gene of interest in a RNA dependent DNA methylation pathway is silenced in the plant;   b) harvesting the seeds; and   c) determining whether the seeds comprise clonal embyros, wherein the presence of seeds comprising clonal embryos indicates the maternal plant can produce parthenogenically-derived clonal embryos.   
     
     
         26 . The method of  claim 25 , wherein the activity of the gene of interest is silenced t using RNA interference. 
     
     
         27 . The method of  claim 25 , wherein the gene of interest is AGO4 (ARGONAUTE 4), AGOG (ARGONAUTE 6), AGO8 (ARGONAUTE 8), AGO9 (ARGONAUTE 9), CMT3 (CHROMOMETHYLASE 3), DCL3 (DICER-LIKE 3), DRM2 (DOMAINS REARRANGED METHYLASE 2), EXS1 (EXTRA SPOROGENOUS CELLS1), IDN2 (INVOLVED IN DE NOVO 2), MET1 (METHYL TRANSFERASE 1), NPRD1a (NUCLEAR POLYMERASE D 1a), NPRD1b (NUCLEAR POLYMERASE D 1b), NPRD2 (NUCLEAR POLYMERASE D 2), NRPE1 (NUCLEAR RNA POLYMERASE E 1), NRPE2 (NUCLEAR RNA POLYMERASE E 2), RDR2 (RNA-DEPENDENT RNA POLYMERASE 2), RDR6 (RNA-DEPENDENT RNA POLYMERASE 6), SGS3 (SUPPRESSOR OF GENE SILENCING 3), SUVH2 (SUPPRESSOR OF VARIEGATION 3-9 HOMOLOG 2), and SUVH9 (SUPPRESSOR OF VARIEGATION 3-9 HOMOLOG 9). 
     
     
         28 . A method of increasing the yield of seeds comprising parthenogenically-derived clonal embryos comprising
 a) obtaining a maternal plant unable to pass on paternally-derived chromosomes to embryos, wherein an activity of a gene of interest in a RNA dependent DNA methylation pathway is silenced in the plant;   b) pollinating the maternal plant;   c) collecting seeds produced by the maternal plant;   d) sorting the seeds comprising parthenogenically-derived clonal embryos from seeds comprising non-clonal embryos.   
     
     
         29 . The method of  claim 28 , wherein sorting seeds comprising parthenogenically-derived clonal embryos from seeds comprising non-clonal embryos is based on phenotype or genotype. 
     
     
         30 . The method of  claim 29 , wherein sorting based on phenotype comprises determining the size, shape, color, or a combination thereof, of the seeds. 
     
     
         31 . The method of  claim 28 , wherein the sorting is performed manually. 
     
     
         32 . The method of  claim 28 , wherein the sorting is performed automatically. 
     
     
         33 . The method of  claim 32 , wherein the automatic sorting comprises a machine comprising an optical detector. 
     
     
         34 . The method of  claim 28 , wherein the sorting is done visually. 
     
     
         35 . The method of  claim 28 , wherein the RNA dependent DNA methylation pathway gene is AGO4 (ARGONAUTE 4), AGO6 (ARGONAUTE 6), AGO8 (ARGONAUTE 8), AGO9 (ARGONAUTE 9), CMT3 (CHROMOMETHYLASE 3), DCL3 (DICER-LIKE 3), DRM2 (DOMAINS REARRANGED METHYLASE 2), EXS1 (EXTRA SPOROGENOUS CELLS1), IDN2 (INVOLVED IN DE NOVO 2), MET1 (METHYL TRANSFERASE 1), NPRD1a (NUCLEAR POLYMERASE D 1a), NPRD1b (NUCLEAR POLYMERASE D 1b), NPRD2 (NUCLEAR POLYMERASE D 2), NRPE1 (NUCLEAR RNA POLYMERASE E 1), NRPE2 (NUCLEAR RNA POLYMERASE E 2), RDR2 (RNA-DEPENDENT RNA POLYMERASE 2), RDR6 (RNA-DEPENDENT RNA POLYMERASE 6), SGS3 (SUPPRESSOR OF GENE SILENCING 3), SUVH2 (SUPPRESSOR OF VARIEGATION 3-9 HOMOLOG 2), and SUVH9 (SUPPRESSOR OF VARIEGATION 3-9 HOMOLOG 9). 
     
     
         36 . A maternal plant comprising a construct, wherein the construct comprises a nucleic acid sequence that renders the maternal plant defective for RNA-dependent DNA methylation. 
     
     
         37 . The maternal plant of  claim 36 , wherein the maternal plant produces seeds comprising parthenogenically-derived clonal embryos. 
     
     
         38 . The maternal plant of  claim 36 , wherein the nucleic acid sequence silences activity of a RNA-dependent DNA methylation pathway gene. 
     
     
         39 . The maternal plant of  claim 36 , wherein the RNA dependent DNA methylation pathway gene is AGO4 (ARGONAUTE 4), AGO6 (ARGONAUTE 6), AGO8 (ARGONAUTE 8), AGO9 (ARGONAUTE 9), CMT3 (CHROMOMETHYLASE 3), DCL3 (DICER-LIKE 3), DRM2 (DOMAINS REARRANGED METHYLASE 2), EXS1 (EXTRA SPOROGENOUS CELLS1), IDN2 (INVOLVED IN DE NOVO 2), MET1 (METHYL TRANSFERASE 1), NPRD1a (NUCLEAR POLYMERASE D 1a), NPRD1b (NUCLEAR POLYMERASE D 1b), NPRD2 (NUCLEAR POLYMERASE D 2), NRPE1 (NUCLEAR RNA POLYMERASE E 1), NRPE2 (NUCLEAR RNA POLYMERASE E 2), RDR2 (RNA-DEPENDENT RNA POLYMERASE 2), RDR6 (RNA-DEPENDENT RNA POLYMERASE 6), SGS3 (SUPPRESSOR OF GENE SILENCING 3), SUVH2 (SUPPRESSOR OF VARIEGATION 3-9 HOMOLOG 2), and SUVH9 (SUPPRESSOR OF VARIEGATION 3-9 HOMOLOG 9). 
     
     
         40 . A seed comprising a parthenogenically-derived clonal embryo comprising a defective RNA-dependent DNA methylation pathway gene. 
     
     
         41 . The seed of  claim 40 , wherein the seed, when grown, produces seeds comprising clonal embryos. 
     
     
         42 . The seed of  claim 40 , wherein the RNA dependent DNA methylation pathway gene is AGO4 (ARGONAUTE 4), AGOG (ARGONAUTE 6), AGO8 (ARGONAUTE 8), AGO9 (ARGONAUTE 9), CMT3 (CHROMOMETHYLASE 3), DCL3 (DICER-LIKE 3), DRM2 (DOMAINS REARRANGED METHYLASE 2), EXS1 (EXTRA SPOROGENOUS CELLS1), IDN2 (INVOLVED IN DE NOVO 2), MET1 (METHYL TRANSFERASE 1), NPRD1a (NUCLEAR POLYMERASE D 1a), NPRD1b (NUCLEAR POLYMERASE D 1b), NPRD2 (NUCLEAR POLYMERASE D 2), NRPE1 (NUCLEAR RNA POLYMERASE E 1), NRPE2 (NUCLEAR RNA POLYMERASE E 2), RDR2 (RNA-DEPENDENT RNA POLYMERASE 2), RDR6 (RNA-DEPENDENT RNA POLYMERASE 6), SGS3 (SUPPRESSOR OF GENE SILENCING 3), SUVH2 (SUPPRESSOR OF VARIEGATION 3-9 HOMOLOG 2), and SUVH9 (SUPPRESSOR OF VARIEGATION 3-9 HOMOLOG 9).

Join the waitlist — get patent alerts

Track US2016208282A1 — get alerts on status changes and closely related new filings.

We store only your email — no account needed. See our privacy policy.